CoChat
Sign Up
Cited by 376Telomeres, Telomerase, and SenescenceGenetics, Aging, and Longevity in Model OrganismsSingle-cell and spatial transcriptomics

Guidelines for minimal information on cellular senescence experimentation in vivo

Open access
Ogrodnik, Mikolaj, Carlos Acosta, Juan, Adams, Peter D., d’Adda di Fagagna, Fabrizio, Baker, Darren J., Bishop, Cleo L., Chandra, Tamir, Collado, Manuel +24 more·2024·Cell
DOI 10.1016/j.cell.2024.05.059PMC PMC11790242
Open this paper in CoChat — free accountOpen PDF

Abstract

Cellular senescence is a cell fate triggered in response to stress and is characterized by stable cell-cycle arrest and a hypersecretory state. It has diverse biological roles, ranging from tissue repair to chronic disease. The development of new tools to study senescence in vivo has paved the way for uncovering its physiological and pathological roles and testing senescent cells as a therapeutic target. However, the lack of specific and broadly applicable markers makes it difficult to identify and characterize senescent cells in tissues and living organisms. To address this, we provide practical guidelines called "minimum information for cellular senescence experimentation in vivo" (MICSE). It presents an overview of senescence markers in rodent tissues, transgenic models, non-mammalian systems, human tissues, and tumors and their use in the identification and specification of senescent cells. These guidelines provide a uniform, state-of-the-art, and accessible toolset to improve our understanding of cellular senescence in vivo.

Integrity signals

Registry

Crossref · 10.1016/j.cell.2024.05.059

Cites

169 of 169 references verified0 citation errors · 0 not checkable

Statistics

Nothing to recompute.

Claims the paper makes

182 of 236 cited sources checked · 15 supported · 1 flagged · accuracy not yet measured

  • Premisecitation

    The establishment of minimum-information guidelines such as MIAME and MISEV improved the quality, harmonization, and reproducibility of data reporting.

    “The establishment of the so-called “minimum information for experimentation” (for example, “Minimum information about a microarray experiment” (MIAME) or “Minimal information for studies of extracellular vesicles” (MISEV)) led to an improvement in data reporting quality, harmonization, and reproducibility2,3.”
    Quote in the paperSubstantial claim1 of 2 sources checked
    • cites [2] source

      The Introduction identifies MISEV as minimum-information guidelines intended to improve EV reporting, reliability, and reproducibility. But it says the MISEV2014 authors *hoped* to improve reliability and reproducibility; it does not establish that the guidelines led to the claimed improvements in reporting quality, harmonization, and reproducibility. This source concerns MISEV, while the sentence also cites another source for MIAME. (LLM, conf=0.85)

      Experimental — accuracy not yet measured
    • cites [3] source
  • Premisecitation

    Persistent activation of the DNA damage response contributes to stable cell-cycle arrest and transcriptional induction of pro-inflammatory SASP factors.

    “This leads to persistent activation of the DNA damage response (DDR), which contributes to stable cell-cycle arrest and transcriptional induction of pro-inflammatory SASP factors16–18.”
    Quote in the paperNot a substantial claim2 of 3 sources checked
    • cites [16] source

      The Introduction (page not provided) describes senescence as irreversible cell-cycle arrest and says senescent cells secrete inflammatory SASP factors. The Results section on ALISE and the SASP (page not provided) reports DNA-damage foci and SASP components, but does not establish that persistent DDR activation causes both arrest and transcriptional induction of those factors; SASP components declined after lipid deprivation even though DNA-damage foci did not. (LLM, conf=0.84)

      Experimental — accuracy not yet measured
    • cites [17] source
    • cites [18] source
  • Premisecitation

    In vivo, some cells such as macrophages may have inherently high β-galactosidase activity due to high lysosomal mass and/or number.

    “In addition, in vivo, a number of cells, such as macrophages, might have an inherently high β-galactosidase activity due to the high mass and/or number of lysosomes44.”
    Quote in the paperSubstantial claim1 of 1 sources checked
    • cites [44] source

      The Materials and Methods sections on the SA-β-Gal assay and whole-mount SA-β-Gal staining describe staining procedures, but do not discuss inherently high β-galactosidase activity in macrophages or attribute it to lysosomal mass or number. Searches for macrophages and lysosomes found no passage addressing that explanation; page numbers were unavailable in our copy. (LLM, conf=0.76)

      Experimental — accuracy not yet measured
  • Premisecitation

    Several recent publications have successfully used anti-murine p16Ink4a antibodies in various applications.

    “Nonetheless, several recent publications have successfully used anti-murine p16Ink4a antibodies in various applications6–8.”
    Quote in the paperNot a substantial claim2 of 3 sources checked
    • cites [6] source

      The Key Resources Table (Additional information; page not provided) lists a p21 antibody but no anti-murine p16Ink4a antibody. Searches for p16Ink4a, Cdkn2a, and anti-murine p16 antibodies found no passage describing their use, so our copy cannot verify the attributed claim. (LLM, conf=0.78)

      Experimental — accuracy not yet measured
    • cites [7] source
    • cites [8] source
  • Premisecitation

    SA-β-gal reflects the activity of a lysosomal enzyme that cleaves terminal β-D-galactose residues from β-D-galactosides.

    “One of the most recognizable and common markers of cellular senescence is senescence-associated beta-galactosidase (SA-β-gal)37, which reflects the activity of a lysosomal enzyme that cleaves terminal β-d-galactose residues from β-D-galactosides38.”
    Quote in the paperNot a substantial claim2 of 2 sources checked
    • cites [37] source

      The Results section reports high SA-β-gal activity as evidence of senescence, but does not explain the enzyme’s location or what it cleaves. Searches for lysosomal β-galactosidase and terminal galactose cleavage found no passage addressing the biochemical claim attached to this citation; page numbers were not available in our copy. (LLM, conf=0.75)

      Experimental — accuracy not yet measured
    • cites [38] source

      The Results passages concern transcription and DNA-damage responses, not SA-β-gal or its enzymatic activity. Searches for SA-β-gal, lysosomal enzyme, galactose, and galactosides found no passage in our copy addressing the claimed cleavage reaction; the References section does not establish it. (LLM, conf=0.76)

      Experimental — accuracy not yet measured
  • May combine two claims. 67
  • Classified as probably not a substantial claim. 25
  • Not yet classified. 1

  • May combine two claims.

    Premisecitation

    p16 is often used as a prognostic marker in some cancers, but in cancers it rarely induces cell cycle arrest.

    “Indeed, p16Ink4a is often used as a prognostic marker in some cancers, such as breast, colon, or head and neck cancer; however, in cancers, p16 rarely induces cell cycle arrest151.”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [151] source

      The Results state that p16INK4a acts to promote cell-cycle arrest, and the Discussion reports that disrupting INK4a disabled drug-induced arrest in the lymphoma model. That conflicts with the claim that p16 rarely induces arrest in cancers. The paper links INK4a status to treatment outcome, but the passages do not establish its use as a prognostic marker in breast, colon, or head and neck cancer; no page numbers are available in our copy. (LLM, conf=0.84)

      Checked by a reasoning model — may be wrong.

      Experimental — accuracy not yet measured

  • May combine two claims.

    Premisecitation

    Additional cyclin-dependent kinase inhibitors such as p15Ink4b, p19Ink4d, and p27Kip1 may participate in senescence induction, but current evidence is insufficient to use their expression as in situ or in vivo senescence markers.

    “Additional CDKIs, such as p15Ink4b, p19Ink4d, and p27Kip1, could be potentially involved in the induction of cellular senescence5, but there is currently little evidence to support the use of their expression as markers of senescence in situ or in vivo.”
    Quote in the paperSubstantial claim1 of 1 sources checked
    • cites [5] source

      The citation follows the sentence’s claim that p15Ink4b, p19Ink4d, and p27Kip1 may help induce senescence; the subsequent statement about their suitability as markers is not attached to it. In Results (“Perforin deficiency accelerates senescence with age”; page unavailable), the paper reports p15 expression among markers examined in senescent tissues, but does not establish a role in induction or address p19Ink4d or p27Kip1. (LLM, conf=0.78)

      Experimental — accuracy not yet measured

  • May combine two claims.

    Premisecitation

    γ-H2A.X is formed by post-translational modifications triggered by double-strand breaks, while 53BP1 is recruited to damage sites through RNA-mediated condensation.

    “γ-H2A.X is formed by post-translational modifications triggered by DSBs, while 53BP1 is recruited to DSBs in response to damage undergoing RNA-mediated condensation21.”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [21] source

      The Results section on p16-expressing FAPs describes γH2A.X-positive nuclei as a mark of DNA damage, and the Methods section describes staining for γH2A.X. Neither passage explains how γH2A.X forms. Searches for double-strand-break-triggered modification and RNA-mediated recruitment of 53BP1 found no passage addressing either mechanism. (LLM, conf=0.76)

      Experimental — accuracy not yet measured

  • May combine two claims.

    Premisecitation

    The CDKN2A locus produces another mRNA encoding p19Arf in mouse and p14ARF in human, which shares the second exon with p16Ink4a and creates sequence similarity between the transcripts.

    “One consideration for measuring p16Ink4a mRNA is that the CDKN2A locus encodes another mRNA coding for p19Arf (mouse)/p14ARF (human), which shares the second exon of CDKN2A, generating a region of sequence similarity between these two transcripts5.”
    Quote in the paperSubstantial claim1 of 1 sources checked
    • cites [5] source

      The Results section, ‘Perforin deficiency accelerates senescence with age’ (page unavailable), identifies p16 with Cdkn2a and measures p16 expression. It does not address an ARF transcript, a shared second exon, or sequence similarity. Searches for p19Arf/p14ARF and CDKN2A alternative transcripts found no passage establishing the claim. (LLM, conf=0.73)

      Experimental — accuracy not yet measured

  • May combine two claims.

    Premisecitation

    Release of HMGB1 from the nucleus is an early sign of senescence induction and can be detected in situ.

    “The release of HMGB1 from the nucleus is an early sign of senescence induction13 and can be detected in situ (Fig. 1 D).”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [13] source

      The Results section discusses telomeric DNA-damage foci during senescence, while the Methods > Immunofluorescence section lists antibodies for Ki67, γH2A.X and 53BP1. Searches for HMGB1 and nuclear release found no passage addressing whether HMGB1 release is an early sign of senescence. The in situ detection statement points to the citing paper’s own Fig. 1 D, not explicitly to this source. (LLM, conf=0.76)

      Experimental — accuracy not yet measured

  • May combine two claims.

    Premisecitation

    PCNA and KI67 levels estimate cell cycle phase, with PCNA highest in S phase and KI67 highest in M phase.

    “Specifically, the levels of PCNA and KI67 provide an estimation of which cell cycle phase a cell is in, with PCNA being at the highest level in the S phase and KI67 in the M phase9,10 (Fig. 1 B).”
    Quote in the paperNot a substantial claim2 of 2 sources checked
    • cites [9] source

      The Results section on senescent stellate cells calls Ki-67 a proliferation-associated marker, and Experimental procedures lists a Ki67 antibody. Neither passage addresses whether its level identifies a cell-cycle phase. Searches for PCNA and for the claimed S- and M-phase peaks found no supporting passage in our copy; page numbers were not provided. (LLM, conf=0.72)

      Experimental — accuracy not yet measured
    • cites [10] source

      The Methods section ('Antibodies for IHC/IF'; page unavailable) lists antibodies for PCNA and Ki-67, and the Results use PCNA-negative cells as a senescence marker. Searches for cell-cycle phases and peak expression found no passage saying that their levels estimate a cell’s phase, that PCNA peaks in S phase, or that Ki-67 peaks in M phase. (LLM, conf=0.76)

      Experimental — accuracy not yet measured

  • May combine two claims.

    Interpretationreasoning

    Generic probes and primers designed for CDKN2A transcripts can detect both Arf and Ink4a mRNAs, so detection strategies must be validated.

    “Thus, certain generic probes and primers designed to detect transcripts from the CDKN2A locus can simultaneously measure Arf and Ink4a mRNAs, making the validation of detection strategies essential.”
    Quote in the paperSubstantial claim1 of 1 sources checked
    • cites [5] source

      The Methods (Quantitative PCR; page unavailable) describe measuring mRNA and refer primer sequences to supplementary methods. The Results (Perforin deficiency accelerates senescence with age; page unavailable) report p16 expression, but neither passage addresses whether CDKN2A probes or primers detect both Arf and Ink4a, or whether detection strategies require validation. Searches for those terms and for primer specificity found no decisive passage in our copy. (LLM, conf=0.76)

      Experimental — accuracy not yet measured

  • May combine two claims.

    Premisecitation

    HMGB1 loss or release is triggered by various stress conditions including necrosis and apoptosis, making it unfeasible as a sole senescence marker.

    “In addition, HMGB1 loss/release is triggered under various stress conditions, including necrosis and apoptosis14, making its use as a sole marker of senescence unfeasible.”
    Quote in the paperSubstantial claim1 of 1 sources checked
    • cites [14] source

      The Results section on dietary restriction discusses TAF, SADS and karyomegaly as senescence markers, but does not discuss HMGB1. Searches for HMGB1 loss or release, necrosis, apoptosis, and its suitability as a sole marker found no passage in our copy addressing the claim. (LLM, conf=0.74)

      Experimental — accuracy not yet measured

  • May combine two claims.

    Premisecitation

    Telomeric DNA damage is more senescence-specific and can be quantified by combining FISH for telomeric sequences with immunostaining for DNA damage response proteins.

    “Telomeric DNA damage is more senescence-specific and can be quantified by combining fluorescence in situ hybridization (FISH) for telomeric sequences and immunostaining (immunoFISH) for DSB-associated proteins22.”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [22] source

      The Methods section on immunofluorescence and quantification describes staining and counting DNA-damage markers, but not telomeric FISH or immunoFISH. Searches for telomeric DNA damage and telomere-associated damage foci found no passage in our copy establishing that the measure is more senescence-specific or describing the claimed combined assay. Page numbers were not available. (LLM, conf=0.76)

      Experimental — accuracy not yet measured

  • May combine two claims.

    Premisecitation

    Lipofuscin, composed of heavily oxidized proteins/lipoproteins and metals, resists lysosomal hydrolysis, forms intra-lysosomal aggregates, and eventually causes dysfunctional lysosomes and impaired autophagic flux.

    “Lipofuscin, comprising heavily oxidized proteins/lipoproteins and metals, is resistant to hydrolysis by lysosomal enzymes and forms intra-lysosomal aggregates, eventually generating dysfunctional lysosomes and impaired autophagic flux33.”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [33] source

      In “Importance of protein and lipid damage in senescent cells” (page unavailable), the paper describes lipofuscin as a nondegradable product of protein and lipid oxidation that resides primarily in lysosomes. The same section reports impaired autophagic flux in senescent cells, but does not establish that lipofuscin causes it through intra-lysosomal aggregation and lysosomal dysfunction; it also does not identify metals as a component. (LLM, conf=0.88)

      Experimental — accuracy not yet measured

  • May combine two claims.

    Premisecitation

    Decondensation occurs in essentially all centromeric satellites in vitro, but only a fraction of centromeric satellites undergo unwinding in vivo.

    “Moreover, while decondensation occurs in essentially all centromeric satellites in vitro24, only a fraction of centromeric satellites undergo unwinding in vivo25,26 (Fig. 1 F).”
    Quote in the paperSubstantial claim2 of 3 sources checked
    • cites [24] source
    • cites [25] source

      The Introduction and Results discuss senescent cells in the murine skeleton, not centromeric satellites. Searches for centromeric-satellite unwinding and decondensation found no passage addressing the claimed in vitro–in vivo difference. (LLM, conf=0.76)

      Experimental — accuracy not yet measured
    • cites [26] source

      The STAR METHODS passages describe telomere staining and other assays in adipose tissue, not centromeric satellites. Searches for centromeric-satellite unwinding in vivo and decondensation in vitro found no passage addressing the claim; page numbers were not supplied. (LLM, conf=0.78)

      Experimental — accuracy not yet measured

  • May combine two claims.

    Premisecitation

    Secreted SASP factors are normally small proteins poorly retained within cells, making IHC/IHF detection highly challenging, although some success has been reported for IL-1α and IL-1β.

    “The origin of SASP factors can be traced using histological methods; however, secreted factors are normally small proteins poorly retained within cells, which makes their detection using IHC/IHF highly challenging, although there has been some success, for example, with IHC against IL-1α29 and IL-1β30.”
    Quote in the paperSubstantial claim2 of 2 sources checked
    • cites [29] source

      Section 3 discusses p21 regulation during the cell cycle, and Section 2 discusses p21 binding to PCNA; neither addresses detection of secreted SASP factors. Searches for SASP, IHC/IHF, and IL-1α found no passage reporting successful IL-1α immunohistochemistry, the specific point to which this citation is attached. (LLM, conf=0.78)

      Experimental — accuracy not yet measured
    • cites [30] source

      The Introduction describes the SASP as secretion of cytokines, growth factors, and proteases. The Immunofluorescence methods describe staining for lamins, not IL-1α or IL-1β. Searches for IL-1 staining and poor cellular retention found no passage addressing the claimed difficulty of IHC/IHF detection or the reported successes with either interleukin. (LLM, conf=0.76)

      Experimental — accuracy not yet measured

  • May combine two claims.

    Premisecitation

    SASP inducers are diverse, but in most cases they depend on NF-κB, CEBPβ, and/or STAT signaling.

    “SASP inducers are diverse; however, in most cases, they rely on the involvement of NF-κB, CEBPβ, and/or STAT signaling28.”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [28] source

      The Introduction and section 3 discuss Ki-67 in cell-cycle regulation and mention senescent cells, but neither addresses SASP induction. Searches for SASP and for NF-κB, CEBPβ, and STAT signaling found no passage in our copy supporting the attributed claim. (LLM, conf=0.76)

      Experimental — accuracy not yet measured

  • May combine two claims.

    Premisecitation

    The senescence-associated secretory phenotype is a core feature of cellular senescence and a major mediator of the pathophysiological functions of senescent cells.

    “Increased production and secretion of cytokines, growth factors, matrix-remodeling enzymes, lipids, extracellular vesicles, and miRNAs, generally defined as the senescence-associated secretory phenotype (SASP)17,27, is a core feature of cellular senescence and a major mediator of the pathophysiological functions of senescent cells.”
    Quote in the paperNot a substantial claim1 of 2 sources checked
    • cites [17] source

      The Introduction (page unavailable) describes a pro-inflammatory secretion pattern as a major change in senescent cells, supporting the claim that a secretory phenotype is a core feature. The Discussion (page unavailable) says inflammatory stimuli from senescent cells might contribute to bystander effects and age-related loss of organ function, but it does not establish SASP as a major mediator of senescent cells’ pathophysiological functions generally. (LLM, conf=0.84)

      Experimental — accuracy not yet measured
    • cites [27] source

  • May combine two claims.

    Premisecitation

    With aging, there is a marked increase in CDKN2A mRNA levels in osteocyte-enriched bone samples, with a modest increase in CDKN1A mRNA levels.

    “With aging, there was a marked increase in CDKN2A mRNA levels in the osteocyte-enriched bone samples (Fig. 2 D), with a modest increase in CDKN1A mRNA levels73.”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [73] source

      The Results passage on Fig. 2D compares gene-expression profiles of young and aged skin stem cells, not osteocyte-enriched bone samples. Searches for CDKN2A, CDKN1A, and osteocytes found no passage reporting the claimed mRNA changes. (LLM, conf=0.76)

      Experimental — accuracy not yet measured

  • May combine two claims.

    Premisecitation

    Senescent cells accumulate with aging and skeletal injury in the bone microenvironment.

    “There is considerable evidence that senescent cells accumulate with aging and skeletal injury in the bone microenvironment25,73–76.”
    Quote in the paperNot a substantial claim5 of 5 sources checked
    • cites [25] source

      The Introduction states that senescent cells accumulate in the bone microenvironment with age, and the Results report increased percentages of senescent skeletal cells in aged mice. The Discussion links injury to a possible role for p21-positive senescent cells and reports p21 upregulation after bone fracture, but does not establish that senescent cells accumulate in the bone microenvironment following skeletal injury. (LLM, conf=0.84)

      Experimental — accuracy not yet measured
    • cites [73] source

      The Abstract and Discussion describe aging hair-follicle stem cells, the skin microenvironment, and impaired hair regeneration after skin injury. Searches for senescent cells, bone, and skeletal injury found no passage in our copy addressing accumulation of senescent cells in the bone microenvironment. (LLM, conf=0.78)

      Experimental — accuracy not yet measured
    • cites [74] source

      The paper states that cells bearing senescence markers accumulate in tissues with age (opening background passage, indexed under Abstract; page unavailable). Its study concerns lung aging and COPD, not the bone microenvironment or skeletal injury, so those parts of the cited claim are not supported. (LLM, conf=0.88)

      Experimental — accuracy not yet measured
    • cites [75] source

      The Article section says cellular senescence occurs in ageing and wound healing. It does not say that senescent cells accumulate after skeletal injury or locate that accumulation in the bone microenvironment; searches for skeletal injury, bone, and fracture found no supporting passage. Page numbers were not available in our copy. (LLM, conf=0.78)

      Experimental — accuracy not yet measured
    • cites [76] source

      The Results section (“Bleomycin injury induces cell senescence”; page unavailable) reports a senescent signature after lung injury, and the paper discusses senescence in the context of ageing. It does not establish that senescent cells accumulate with ageing or skeletal injury in the bone microenvironment; those are different conditions and tissue. (LLM, conf=0.83)

      Experimental — accuracy not yet measured

  • May combine two claims.

    Premisecitation

    p16+ cells may predominate in aged adipose tissue, whereas p21 predominates in obesity.

    “For example, p16+ cells may predominate in aged adipose tissue, whereas p21 predominates in obesity84,85.”
    Quote in the paperNot a substantial claim2 of 2 sources checked
    • cites [84] source

      The Introduction discusses p16 and p21 in aging bone marrow, not aged adipose tissue. Searches for aged adipose tissue and obesity with p21 found no passage comparing which marker predominates in those conditions; the obesity-related material appears only in the References. (LLM, conf=0.75)

      Experimental — accuracy not yet measured
    • cites [85] source

      The Discussion (page unavailable) contrasts p16-positive cells in age-related bone loss with p21-positive cells in radiation-induced bone loss and marrow adiposity. Searches for aged adipose tissue and obesity found no passage comparing which cell population predominates in those conditions. (LLM, conf=0.74)

      Experimental — accuracy not yet measured

  • May combine two claims.

    Premisecitation

    In aged vs. young mouse bones, there is an increase in p16+, KI67−, BCL2+, and SASP+ cells (and an analogous increase in p21+ population) in late osteoblast/osteocytic and bone mesenchymal cells.

    “In aged vs. young mouse bones, there is an increase in p16+, KI67−, BCL2+, and SASP+ cells (and an analogous increase in the p21+ population) in late osteoblast/osteocytic and bone mesenchymal cells6.”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [6] source

      The sections “The zone encompasses healing processes” and “P-rpS6 deficiency accelerates wound closure” discuss p21-positive keratinocytes in wounded skin, not aged versus young mouse bones. Searches for osteoblasts, osteocytes, bone mesenchymal cells, and the specified marker combination found no passage addressing the claimed age-related increases. Page numbers were not available in our copy. (LLM, conf=0.78)

      Experimental — accuracy not yet measured

  • May combine two claims.

    Premisecitation

    The outer layer of the animal body is readily accessible for sample collection and is suitable as an index organ for aging and age-related pathologies of internal organs.

    “The outer layer of the animal body is readily accessible for sample collection and is suitable as an index organ for aging and age-related pathologies of internal organs, such as the cardiovascular system, bones, and brain51–53, among others.”
    Quote in the paperSubstantial claim2 of 3 sources checked
    • cites [51] source

      The Introduction discusses senescence markers in aging tissues, and Experimental procedures describes fibroblasts obtained from skin biopsies. Neither section says skin is readily accessible for sampling or can serve as an index of aging and disease in internal organs. Searches for those concepts returned no supporting passage; page numbers were not available in our copy. (LLM, conf=0.76)

      Experimental — accuracy not yet measured
    • cites [52] source
    • cites [53] source

      The Introduction links cellular senescence to age-related disorders, and the Histology section describes tissue staining, but neither identifies skin as an accessible sampling site or an index of aging and disease in internal organs. Searches for skin, skin biopsy, and an index organ found no passage addressing that claim. (LLM, conf=0.75)

      Experimental — accuracy not yet measured

  • May combine two claims.

    Premisecitation

    SA-β-gal is more intense in senescence-high conditions such as aging, but is also detectable in neurons of young and healthy mice.

    “certain senescence markers, such as SA-β-gal, despite being more intense in senescence-high conditions, such as aging, are also detectable in the neurons of young and healthy mice46,50”
    Quote in the paperSubstantial claim1 of 2 sources checked
    • cites [46] source

      The Deregulated Metabolic Profile section describes SA-β-gal as prominent in senescent cells, and Murine Models of Accelerated Senescence and Aging reports SA-β-gal-positive cells in mice with accelerated aging. The copy does not establish that staining is more intense with ordinary aging or that SA-β-gal is detectable in neurons of young, healthy mice. (LLM, conf=0.83)

      Experimental — accuracy not yet measured
    • cites [50] source

  • May combine two claims.

    Premisecitation

    In animal models of IPF, bleomycin-induced injury and telomere deficiency drive fibroblasts and epithelial cells towards increased senescence, a feature also observed in lung tissue of IPF patients.

    “In animal models of idiopathic pulmonary fibrosis (IPF), bleomycin-induced injury66 and telomere deficiency72 drive fibroblasts and epithelial cells towards increased levels of senescence, a feature also observed in the lung tissue of patients with IPF.”
    Quote in the paperNot a substantial claim1 of 2 sources checked
    • cites [66] source
    • cites [72] source

      The Results section on aged keratinocytes addresses wound repair in mouse skin, not senescence in pulmonary fibrosis. Searches for bleomycin, telomere deficiency, and lung fibrosis found no passage in our copy supporting the claim attributed to citation 72. (LLM, conf=0.77)

      Experimental — accuracy not yet measured

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    The lung can be driven into pathological conditions by external stimuli including smoke exposure and viral infections.

    “The lung can be driven into pathological conditions by a variety of external stimuli, including smoke exposure64 and viral infections65,71.”
    Quote in the paperNot a substantial claim2 of 3 sources checked
    • cites [64] source
    • cites [65] source

      The Introduction mentions chronic lung disease among pathologies associated with senescent cells, and Results > The p16-3MR Mouse Model reports a response to irradiation in mouse lungs. Neither passage links lung pathology to smoke exposure or viral infection; searches for those stimuli found no supporting passage in our copy. Page numbers were not available. (LLM, conf=0.74)

      Experimental — accuracy not yet measured
    • cites [71] source

      The Introduction discusses fluorescence imaging, and Methodology discusses imaging mice; neither addresses lung pathology. Searches for smoke exposure and viral infection found no passage linking either to pathological lung conditions. In the citing sentence, this source appears to be cited for viral infections, but our copy does not address that point. (LLM, conf=0.76)

      Experimental — accuracy not yet measured

  • May combine two claims.

    Premisecitation

    Fibroblasts present papillary and reticular phenotypes and heterogeneous populations at the single-cell level.

    “fibroblasts present not only papillary and reticular phenotypes, but also heterogeneous populations if studied at the single-cell level58,59.”
    Quote in the paperNot a substantial claim2 of 2 sources checked
    • cites [58] source

      The cited paper’s Materials and Methods passages describe rat hippocampal tissue, dendritic spines, and serum assays, not fibroblast phenotypes. Searches for papillary and reticular fibroblasts and for single-cell fibroblast heterogeneity found no passage addressing the claim. No page numbers were provided for these passages. (LLM, conf=0.76)

      Experimental — accuracy not yet measured
    • cites [59] source

      The Methods (‘Single cell preparation and FACS’) and Extended Data Figures 4–5 describe sorting and identifying brain cell populations, not fibroblast populations. Searches for papillary and reticular fibroblasts and single-cell fibroblast heterogeneity found no passage addressing either part of the claim. (LLM, conf=0.78)

      Experimental — accuracy not yet measured

  • May combine two claims.

    Premisecitation

    There are numerous distinct cell types within the lung required for functional substructures, and there is heterogeneity of senescence phenotypes across lung cell types.

    “Like many other tissues, there are numerous distinct cell types within the lung required for creating functional substructures; thus, there is also heterogeneity of senescence phenotypes across lung cell types64–70 (Fig. 2 C).”
    Quote in the paperSubstantial claim3 of 7 sources checked
    • cites [64] source
    • cites [65] source

      The Results section on senescent endothelial and mesenchymal cells reports that fibroblasts and endothelial cells, but not epidermal cells, predominated among senescent cells after skin injury. This supports cell-type differences in senescence in skin, but the paper does not establish the claimed heterogeneity across lung cell types or discuss their functional substructures. (LLM, conf=0.83)

      Experimental — accuracy not yet measured
    • cites [66] source
    • cites [67] source

      The Results and Discussion describe distinct cell populations and stem-cell niches in skin, with senescent fibroblasts disrupting those niches. This supports skin as an example of tissue complexity, but the paper does not establish heterogeneity of senescence phenotypes across lung cell types; searches for lung cell types and lung senescence returned only skin-focused passages. (LLM, conf=0.78)

      Experimental — accuracy not yet measured
    • cites [68] source

      The Results sections on fibroblast subpopulations and aging describe functionally distinct cell populations in human skin and differences among aging dermal fibroblast subpopulations (pages not specified). They do not establish that lung cell types have heterogeneous senescence phenotypes or describe their roles in lung substructures. The paper supports the skin-tissue analogy, not its extension to the lung. (LLM, conf=0.78)

      Experimental — accuracy not yet measured
    • cites [69] source
    • cites [70] source

  • May combine two claims.

    Premisecitation

    Age-related changes in murine liver cells include increased SA-β-gal activity, DNA damage, TAFs, karyomegaly, SADS, ALISE, LINE-1 elements, and SASP factors, some specified to occur in hepatocytes.

    “Age-related changes in murine liver cells include increased SA-β-gal activity78, DNA damage, TAFs19,20,78, karyomegaly, SADS26, ALISE26, LINE-1 elements79, and SASP factors40, some of them specified to occur in hepatocytes19,26.”
    Quote in the paperNot a substantial claim3 of 3 sources checked
    • cites [78] source

      The aged-mouse-lung analysis reports SASP factors in age-associated macrophages, supporting the SASP portion attached to this citation. It does not establish increased SASP factors in murine liver cells or hepatocytes; the other listed changes have separate citations and are not attributed to this source. (LLM, conf=0.86)

      Experimental — accuracy not yet measured
    • cites [19] source

      The Abstract (page unavailable) says senescent cells secrete bioactive molecules, including interleukins and growth factors, and reports senescence-marker-positive hepatocytes in mouse livers. It does not show that SASP factors increase with age in murine liver cells. The other listed markers have separate citations in the citing sentence and are not attributed to this paper. (LLM, conf=0.85)

      Experimental — accuracy not yet measured
    • cites [20] source

      The passage on plaque-rich aortic arches (section labeled Abstract; page unavailable) reports elevated SASP components in mice, supporting the SASP-factors portion attached to this citation. It concerns atherosclerotic tissue, not age-related changes in murine liver cells or hepatocytes; the other listed markers are attached to different citations. (LLM, conf=0.87)

      Experimental — accuracy not yet measured

  • May combine two claims.

    Premisecitation

    Elimination of senescent cells alleviates age-related pathologies in BubR1 and Ercc1 mutant mice, but does not restore their lifespan to that of wild-type animals.

    “For example, while the elimination of senescent cells alleviates age-related pathologies in BubR1 and Ercc1 mutant mice, it does not restore their lifespan to that of wild-type animals86,102.”
    Quote in the paperSubstantial claim1 of 2 sources checked
    • cites [86] source
    • cites [102] source

      The Introduction (page unavailable) reports that inactivating p16 partially rescues several age-related phenotypes in a progeroid mouse strain. It does not say that senescent cells were eliminated, address Ercc1 mutant mice, or compare the mutants’ lifespan with that of wild-type animals. (LLM, conf=0.83)

      Experimental — accuracy not yet measured

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Historically, some of the first transgenic animals used for senescence research were knockout models of the major cell cycle inhibitors, p16-KO and p21-KO mice.

    “Historically, some of the first transgenic animals used for senescence research were knockout models of the major cell cycle inhibitors, p16-KO and p21-KO mice97,98.”
    Quote in the paperNot a substantial claim2 of 2 sources checked
    • cites [97] source

      The 'Senescence in T Cells' section (page unavailable) discusses p16INK4a expression and senescence markers in mice, but not p16- or p21-knockout models. Searches for those knockout models and their historical use found no passage establishing that they were among the first transgenic animals used in senescence research. (LLM, conf=0.76)

      Experimental — accuracy not yet measured
    • cites [98] source

      The ‘Cellular senescence and immune cell fate decision’ section (page unavailable) describes knockout mice for the CDKN2A locus, which encodes p16INK4a and p14/p19ARF, and their use in studying senescence. It mentions p21 in the cell-cycle-arrest pathway, but does not identify p21-knockout mice as a research model or say these models were among the first transgenic animals used in senescence research. (LLM, conf=0.86)

      Experimental — accuracy not yet measured

  • May combine two claims.

    Premisecitation

    The promoter activity of p16 is generally weak, and if the neomycin cassette is not removed, its strong promoter may affect the p16 promoter, which could be related to differences observed between studies.

    “Moreover, as mentioned above, the promoter activity of p16 is generally weak, and if the neomycin cassette is not removed, its strong promoter may affect the p16 promoter, which could be related to the differences observed between studies109,110.”
    Quote in the paperNot a substantial claim2 of 2 sources checked
    • cites [109] source

      Section 3 discusses p16-based mouse models, and Section 2 discusses p16 as a senescence marker; neither addresses the strength of the p16 promoter or differences between studies caused by a neomycin cassette. Searches for “neomycin cassette,” “p16 promoter,” and weak promoter activity found no passage supporting the claimed mechanism. Page numbers were not available in our copy. (LLM, conf=0.76)

      Experimental — accuracy not yet measured
    • cites [110] source

      The Materials and Methods and Discussion passages describe a p21 promoter–luciferase reporter, not a p16 promoter. Searches for p16 promoter activity and neomycin-cassette interference found no passage addressing either point or linking them to differences between studies; page numbers were unavailable in our copy. (LLM, conf=0.76)

      Experimental — accuracy not yet measured

  • May combine two claims.

    Premisecitation

    An assay based on analysis of CDKN2A mRNA in PBMCs or selected subsets such as CD3+ T cells was developed and shown to correlate with biological age and exposure to stress such as smoking or diet.

    “To detect the senescence-like state in immune cell populations, an assay based on analysis of CDKN2A mRNA in PBMCs or selected subsets such as CD3+ T cells was developed and shown to correlate with biological age and exposure to stress such as smoking or diet96.”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [96] source

      The Results section reports p16 qPCR in old mouse muscle and senescence markers in mouse hepatocytes, not a CDKN2A mRNA assay in PBMCs or CD3+ T cells. Searches for that assay and for correlations with biological age, smoking, or diet found no passage addressing the claim. (LLM, conf=0.76)

      Experimental — accuracy not yet measured

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Premisecitation

    Age-related changes in the immune system are characterized by altered ratios of naïve and memory T cells, lymphoid to myeloid cells, CD4+ to CD8+ cells, and thymic atrophy.

    “These age-related changes are characterized by altered ratios of naïve and memory T cells, lymphoid to myeloid cells, CD4+ to CD8+ cells, and thymic atrophy.”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [91] source

      The Results section on p16-high and p21-high cells describes immune cell types in aged mouse tissues, but not age-related changes in their ratios. Searches for naïve and memory T cells, CD4/CD8 ratios, lymphoid-to-myeloid ratios, and thymic atrophy found no passage addressing the claimed pattern; the References mention memory-like T cells only in another paper’s title. (LLM, conf=0.76)

      Experimental — accuracy not yet measured

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    With aging, the immune system loses its ability to mount an effective immune response against invading pathogens and cancer cells, termed immunosenescence.

    “With aging, the immune system loses its ability to mount an effective immune response against invading pathogens and cancer cells, a phenomenon termed immunosenescence91.”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [91] source

      The Introduction (page unavailable) discusses cellular senescence and its increase with aging, not an age-related loss of immune responses to pathogens or cancer cells. Searches for “immunosenescence,” immune responses, pathogens, and cancer found no passage defining or substantiating the claimed phenomenon; a reference to impaired immune surveillance concerns senescent-cell accumulation instead. (LLM, conf=0.76)

      Experimental — accuracy not yet measured

  • May combine two claims.

    Premisecitation

    p21 increased with age in a subset of terminally differentiated myofibers, which also displayed other senescence-associated markers such as TAF, SADS, nuclear loss of LMNB1, HMGB1, and a unique SASP.

    “Interestingly, p21 increased with age in a subset of terminally differentiated myofibers, which also displayed other senescence-associated markers such as TAF, SADS, nuclear loss of LMNB1, HMGB1, and a unique SASP88,89.”
    Quote in the paperNot a substantial claim2 of 2 sources checked
    • cites [88] source

      The Discussion (page unavailable) discusses longer-lasting p21 expression in older mouse hepatocytes, not an age-related increase in terminally differentiated myofibers. Searches for myofibers and the specified markers found no passage addressing TAF, SADS, nuclear loss of LMNB1 or HMGB1, or a unique myofiber SASP. (LLM, conf=0.76)

      Experimental — accuracy not yet measured
    • cites [89] source

      The section ‘IL-33 from senescent HSCs promotes obesity-associated HCC development’ reports more p21-positive hepatic stellate cells in high-fat-diet mouse livers, alongside SASP factors. It does not report an age-related increase in p21 in terminally differentiated myofibers or the listed markers in those cells (searched: myofibers, skeletal muscle, TAF, SADS, and HMGB1). (LLM, conf=0.78)

      Experimental — accuracy not yet measured

  • May combine two claims.

    Premisecitation

    FAPs also exhibit pro-inflammatory SASP, TAFs, and SADS.

    “However, FAPs also exhibit pro-inflammatory SASP, TAFs, and SADS.”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [88] source

      The Introduction (page unavailable) says that SASP generally includes pro-inflammatory chemokines and cytokines, supporting the pro-inflammatory SASP component. The paper studies hepatic stellate cells; the passages do not establish that FAPs exhibit this phenotype or address the claimed TAFs and SADS. (LLM, conf=0.78)

      Experimental — accuracy not yet measured

  • May combine two claims.

    Premisecitation

    Adaptive immune function can be improved by short-term treatment with rapamycin, which reduces markers of senescence and SASP in PBMCs.

    “Interestingly, adaptive immune function can be improved by short-term treatment with rapamycin95, which reduces the markers of senescence and SASP in PBMCs93.”
    Quote in the paperNot a substantial claim2 of 2 sources checked
    • cites [95] source

      The Results section reports increased circulating SASP proteins in p21-overexpressing mice, and Materials and Methods describes measuring those proteins in mouse plasma. Neither passage addresses rapamycin treatment or reduced senescence and SASP markers in PBMCs; searches for rapamycin and PBMCs found no supporting passage in our copy. The adaptive-immune-function clause is attached to a different citation. (LLM, conf=0.75)

      Experimental — accuracy not yet measured
    • cites [93] source

      The main text (Fig. 3a–d) reports reduced senescence markers in the adipose tissue, skeletal muscle and eye of AP20187-treated transgenic mice. It does not show that rapamycin reduces senescence or SASP markers in PBMCs. The adaptive-immune-function statement is attached to a different citation (95), so it is not assessed here. (LLM, conf=0.84)

      Experimental — accuracy not yet measured

  • May combine two claims.

    Premisecitation

    Senescent cell identification by scRNAseq has been successful in multiple in vivo studies, providing insights into proper study design.

    “Notably, senescent cell identification by scRNAseq has been successful in multiple in vivo studies, providing valuable insights into proper study design68,129,137,138.”
    Quote in the paperNot a substantial claim3 of 4 sources checked
    • cites [68] source

      The Results section (‘scRNA-seq analysis of sun-protected human skin’; page unavailable) describes an in vivo human-skin scRNA-seq study and explains a design choice intended to limit photoaging as a confounder. The passages retrieved for ‘senescent’ and ‘senescence’ discuss age-related fibroblast changes, but do not report identifying senescent cells. The cited paper therefore supports the study-design aspect, not the attributed success in senescent-cell identification. (LLM, conf=0.82)

      Experimental — accuracy not yet measured
    • cites [129] source
    • cites [137] source

      In “Cellular Senescence and Cellular Aging in Vivo” (page unavailable), the paper reports an in vivo single-cell transcriptomics study that measured p16-positive cells in old mice. It also discusses challenges in designing in vivo senescence assessments, but says scRNA-seq might refine assessment in the future; it does not establish successful senescent-cell identification across multiple in vivo studies or say those studies yielded study-design guidance. (LLM, conf=0.82)

      Experimental — accuracy not yet measured
    • cites [138] source

      The Results section on single-cell heterogeneity describes imaging and mass-spectrometry proteomics, not identification of senescent cells by scRNA-seq. Searches for “senescent,” “senescence,” and “scRNA-seq” found no passage in our copy reporting multiple in vivo senescence studies or lessons for their design; the scRNA-seq mention is a reference to a pancreatic-cancer study. (LLM, conf=0.74)

      Experimental — accuracy not yet measured

  • May combine two claims.

    Premisecitation

    Induction of senescence is a tumor suppressor mechanism and a side effect of several chemotherapeutic drugs.

    “Induction of senescence is a tumor suppressor mechanism143–145 and a side effect of several chemotherapeutic drugs that induce a phenotype that might be considered an acquired premature progeroid syndrome in both normal and cancer cells146.”
    Quote in the paperNot a substantial claim4 of 4 sources checked
    • cites [143] source

      The cited paper’s Abstract discusses senescent cells in aging and their possible physiological roles, but does not address senescence as a tumor-suppressor mechanism or a side effect of chemotherapy. Searches for chemotherapy, tumor suppression, and progeroid effects found no passage supporting the attributed claim. (LLM, conf=0.76)

      Experimental — accuracy not yet measured
    • cites [144] source

      The Introduction describes cellular stress as a trigger of senescence, and the Results discuss senolytic treatment, but neither passage says that senescence is a tumor-suppressor mechanism or a side effect of chemotherapy. Searches for tumor suppression, chemotherapy-induced senescence, and progeroid syndrome found no passage in our copy addressing those claims. (LLM, conf=0.72)

      Experimental — accuracy not yet measured
    • cites [145] source

      The Introduction says senescence is an important physiological defense against malignant transformation, supporting the tumor-suppression part. The paper describes inducing senescence by irradiation in kidney epithelial cells (Results), but does not say that senescence is a side effect of chemotherapeutic drugs or describe a premature progeroid syndrome in normal and cancer cells. (LLM, conf=0.86)

      Experimental — accuracy not yet measured
    • cites [146] source

      The Results and Discussion (page unavailable) describe doxorubicin as a chemotherapeutic agent known to cause senescence and report using it to induce senescence in primary fibroblasts. The paper does not establish the claim about several chemotherapeutic drugs or a premature progeroid phenotype in both normal and cancer cells. Its abstract also links senescence to tumor suppression, although that clause has separate citations in the citing sentence. (LLM, conf=0.88)

      Experimental — accuracy not yet measured

  • May combine two claims.

    Premisecitation

    Reporter transgenic models enable FACS enrichment of p16high and p21high cells and combination with downstream omics analysis.

    “The development of reporter transgenic models has enabled researchers to use FACS to enrich p16high and p21high cells and combine this with downstream omics analysis and single-cell omics transcriptomics (for example,109).”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [109] source

      Section 3 describes transgenic mouse models that permit visualization of p16-expressing cells, and Section 2 identifies p16 and p21 as senescence markers (page numbers unavailable). Neither passage says the models enable FACS enrichment of both p16-high and p21-high cells or links that enrichment to downstream omics analysis. (LLM, conf=0.83)

      Experimental — accuracy not yet measured

  • May combine two claims.

    Premisecitation

    Spatial omics has not yet been used extensively for senescence research but is in development.

    “Spatial omics has not yet been used extensively for senescence research but is in development132–136.”
    Quote in the paperNot a substantial claim5 of 5 sources checked
    • cites [132] source

      The Discussion (page unavailable) proposes developing reagents to study p15 INK4b and ARF in chicken cell senescence, but does not discuss spatial omics. Searches for spatial omics and spatial transcriptomics in senescence research found no passage addressing whether these methods are in development or how extensively they have been used. (LLM, conf=0.76)

      Experimental — accuracy not yet measured
    • cites [133] source

      The Data availability section describes bulk and single-cell RNA-seq and microscopy, while the Short article reports transcriptomic profiling of senescent cells. Searches for spatial omics and spatially resolved transcriptomics found no passage assessing how extensively spatial omics has been used in senescence research or saying it is in development. (LLM, conf=0.73)

      Experimental — accuracy not yet measured
    • cites [134] source

      The Discussion describes imaging senescent tissue regions, and Materials and Methods describes RNA sequencing of flow-sorted lung cells. Neither passage addresses spatial omics or whether its use in senescence research is limited or in development; searches for spatial omics and spatial transcriptomics found no relevant passage. The passages provide no page numbers. (LLM, conf=0.76)

      Experimental — accuracy not yet measured
    • cites [135] source

      The Introduction discusses senescent cells and drug development, while the Discussion describes imaging and transcriptional comparisons in a zebrafish model (page numbers unavailable). Searches for spatial omics and spatial transcriptomics found no passage addressing how extensively spatial omics has been used in senescence research or whether it is in development. (LLM, conf=0.72)

      Experimental — accuracy not yet measured
    • cites [136] source

      The Introduction section on distinct senescence transcriptomes describes single-cell RNA sequencing, and the STAR Methods list imaging separately. Searches for spatial omics, spatial transcriptomics, and spatial methods found no passage assessing how extensively spatial omics has been used in senescence research or saying it is in development. The available passages therefore do not establish the attributed claim. (LLM, conf=0.75)

      Experimental — accuracy not yet measured

  • May combine two claims.

    Premisecitation

    p21 is elevated in salamander cells in regenerating limbs and in the regenerating fins of zebrafish.

    “In contrast, p21 is elevated in salamander cells in regenerating limbs126 and in the regenerating fins of zebrafish118.”
    Quote in the paperNot a substantial claim2 of 2 sources checked
    • cites [126] source

      The Results report elevated p21 in the aged killifish telencephalon, not in regenerating salamander limbs or zebrafish fins. The Discussion mentions injury-induced senescent cells in salamanders and zebrafish but does not say p21 is elevated in those tissues; searches for p21 with both regeneration contexts found no supporting passage. (LLM, conf=0.77)

      Experimental — accuracy not yet measured
    • cites [118] source

      Citation 126 is attached to the salamander-limb claim, not the zebrafish-fin claim. The Methods and Results and Discussion describe human WI-38 fibroblasts and report elevated p21 in those cells, but no passage addresses p21 in regenerating salamander limbs (searched: salamander cells, regenerating limbs). (LLM, conf=0.76)

      Experimental — accuracy not yet measured

  • Not yet classified.

    Premise

    Most conditions that drive senescence, including aging, produce a small number of senescent cells, often estimated at less than 10% based on p16+ cells in situ.

    “Based primarily on the frequency of p16+ cells in situ, most conditions driving senescence, including age, yield a relatively small number of senescent cells, often estimated to be <10%130.”
    Quote in the paperNot yet classified1 of 1 sources checked
    • cites [130] source

      The Experimental Procedures describe counting γ-H2AX-positive kidney cells, and the Discussion describes SA-β-gal staining in zebrafish skin. Neither passage estimates senescent-cell frequency across conditions from p16+ cells in situ. Searches for p16, senescent-cell proportions, and the <10% estimate found no passage addressing the claim. (LLM, conf=0.76)

      Experimental — accuracy not yet measured

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    The recent generation of the p21-GFP transgenic reporter zebrafish is an example of new transgenic models helpful for senescence research.

    “such as the recent generation of the p21-GFP transgenic reporter zebrafish128.”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [128] source

      The Methods describe GFP-labelled fly clones in larval imaginal discs, not a zebrafish reporter. Searches for “p21-GFP transgenic reporter zebrafish” and “zebrafish p21 senescence reporter model” found no passage in our copy addressing the claimed model. (LLM, conf=0.76)

      Experimental — accuracy not yet measured

  • May combine two claims.

    Premisecitation

    Current omics approaches for senescence are limited to unbiased transcriptomics and targeted proteomics on commercial platforms, though other modalities like single-cell proteomics are in development.

    “It is currently limited to unbiased transcriptomics and targeted proteomics on commercial platforms, although other omics modalities are in development (e.g., single-cell proteomics131).”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [131] source

      The Methods sections on RNA isolation and RT-qPCR and on Western blotting describe measurements in the killifish visual system, not the availability or limitations of senescence omics approaches. Searches for unbiased transcriptomics, targeted proteomics, commercial platforms, and single-cell proteomics found no passage addressing the claim. Page numbers were not available in our copy. (LLM, conf=0.76)

      Experimental — accuracy not yet measured

  • May combine two claims.

    Premisecitation

    Pro-survival proteins enable cancer cells to avoid cell death upon acute stress and are targetable for cancer senolysis.

    “Among potential biomarkers, the relevance of pro-survival proteins is that they enable cancer cells to avoid cell death upon acute stress (e.g., during chemotherapy or radiotherapy) and are targetable for cancer senolysis148,149.”
    Quote in the paperNot a substantial claim2 of 2 sources checked
    • cites [148] source

      The Results section (‘Predictors can assist in the characterization and discovery of senotherapies’; page unavailable) identifies BCL-2 family inhibitors as senolytics and reports that they selectively reduced senescent A549 cancer cells. It does not establish that pro-survival proteins enable cancer cells to avoid death during acute stress from chemotherapy or radiotherapy. (LLM, conf=0.85)

      Experimental — accuracy not yet measured
    • cites [149] source

      The Discussion (page unavailable) addresses senescence and cancer, while the Results section describes drug-induced senescence and its detection. Neither states that pro-survival proteins help cancer cells survive acute stress or can be targeted for cancer senolysis; searches for those terms found no supporting passage in our copy. (LLM, conf=0.76)

      Experimental — accuracy not yet measured

  • May combine two claims.

    Premisecitation

    Tumor cells are heterogeneous, with only a fraction positive for senescence markers, except in some mouse models of senescence reactivation.

    “Except in some mouse models of senescence reactivation, such as through p53 restoration153, cells within a tumor are heterogeneous, with only a fraction becoming positive for cellular senescence markers.”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [153] source

      The paper’s References section lists work on tumor senescence, but that listing does not establish how senescence markers are distributed among cells within a tumor. Searches of our copy for tumor heterogeneity, the fraction positive for senescence markers, and p53-restoration mouse models found no passage addressing the claim. (LLM, conf=0.76)

      Experimental — accuracy not yet measured

  • May combine two claims.

    Premisecitation

    p16 overexpression marks highly proliferative HPV-16 positive cervical cancer cells, driven by RB1 inhibition by viral protein E7.

    “For example, p16 overexpression serves as a marker for highly proliferative HPV-16 positive cervical cancer cells, driven by RB1 (pRB) inhibition of the viral protein E7151.”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [151] source

      The Results report high p16INK4a levels in proliferating p53-null lymphomas and increased p16INK4a after chemotherapy, but do not address cervical cancer. Searches for HPV-16, E7, and RB1/pRB inhibition found no passage establishing the claimed marker or mechanism; page numbers were unavailable in our copy. (LLM, conf=0.75)

      Experimental — accuracy not yet measured

  • Probably not a substantial claim: may not pass S4 (based). May combine two claims.

    Interpretationnone given

    The senescent phenotype is heterogeneous, and no single marker is adequate to confirm the state of senescence.

    “The senescent phenotype is heterogeneous and no single marker is adequate to confirm the state of senescence.”
    Quote in the paperNot a substantial claim
    • Rests on an assumption: No single biomarker is sufficient to detect cellular senescence in vivo, and multiplexing remains a necessary requirement.

  • Classified as probably not a substantial claim. May combine two claims.

    Interpretationreasoning

    Oxidative damage is not specific to senescence and is therefore recommended only as an auxiliary marker.

    “Finally, similar to other forms of damage, oxidative damage is not specific to senescence, and is therefore recommended only as an auxiliary marker.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Premisenone given

    Most cells in vivo divide only once every few weeks or months, and many cells are post-mitotic.

    “In contrast, the majority of cells divide in vivo once every few weeks or months, and many cells are in a post-mitotic state.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Interpretationreasoning

    Even in cell culture, SA-β-gal staining gives false-positive results in starved or confluent cells, and its specificity range in vivo and in situ is unknown.

    “Even in cell culture, SA-β-gal staining results in false-positive staining of starved or confluent cells, whereas for in vivo and in situ conditions, the range of specificity of SA-β-gal is unknown.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Interpretationreasoning

    The stability of cell cycle arrest has been canonically linked to cultured senescent cells, but this feature has not yet been verified for senescence in situ or in vivo.

    “Finally, although the stability of cell cycle arrest has been canonically associated with cultured senescent cells, this feature has not yet been verified as being associated with senescence in situ or in vivo.”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim: may not pass S4 (based).

    Interpretationnone given

    Different cell types may be intrinsically resistant or susceptible to senescence depending on the stimuli.

    “Different cell types may be intrinsically resistant or susceptible to senescence (depending on the stimuli); and Different cell types may exhibit different senescence phenotypes (expressing different senescence-associated markers).”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Interpretationnone given

    DNA double-strand breaks are not specific to senescence, and it is currently unclear whether the number of DNA lesions can discriminate senescence in situ.

    “However, DSBs are not specific to senescence, and it is currently unclear whether the number of DNA lesions can be used as a discriminant of senescence in situ.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Interpretationreasoning

    The primary senescence markers are the presence of p21 and/or p16Ink4a, which have guided in vivo senescence research for the last two decades.

    “The first is the presence of p21 and/or p16Ink4a, two markers that have guided research on senescence in vivo in the last two decades.”
    Quote in the paperSubstantial claim
    • Rests on an assumption: p21Cip1/Waf and p16Ink4a are the cyclin-dependent kinase inhibitors most commonly expressed by senescent cells in vivo.

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Interpretationreasoning

    Because 4-HNE provides a relatively small epitope, antibody specificity should be thoroughly established and verified with suitable controls.

    “However, as 4-HNE provides a relatively small epitope, the specificity of the antibody should be thoroughly established and verified using a series of suitable controls (for example, see Table 1).”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Interpretationreasoning

    Current in situ and in vivo methods provide only a snapshot of cell physiology, and except for transgenic tracker animals, persistence of p16Ink4a or p21Cip1/Waf expression cannot yet be validated.

    “The available methodology for in situ and in vivo measurements offers only a snapshot of cell physiology and, apart from transgenic “tracker” animals, it is not yet possible to validate the persistence of p16Ink4a or p21Cip1/Waf expression in situ or in vivo.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Premisecitation

    Senescence-associated decondensation of satellites is a senescence-associated chromatin change with an established method for in situ detection.

    “One feature of senescence-associated changes in chromatin with an established methodology for in situ detection is senescence-associated decondensation of satellites24 (SADS; Fig. 1 F).”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [24] source

  • Probably not a substantial claim: may not pass S4 (based). May combine two claims.

    Interpretationnone given

    Different cell types may exhibit different senescence phenotypes, expressing different senescence-associated markers.

    “Different cell types may exhibit different senescence phenotypes (expressing different senescence-associated markers).”
    Quote in the paperNot a substantial claim

  • Classified as probably not a substantial claim. May combine two claims.

    Interpretationreasoning

    The extent to which SASP transcript levels reflect protein levels is not well established, so caution is advised when drawing conclusions from such quantitative analyses.

    “However, as with all approaches of RNA measurements, the extent to which the transcript level reflects the protein level is not well established; thus, caution is advised when drawing conclusions from these quantitative analyses.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Interpretationreasoning

    Loss of proliferation markers is a strong indicator of senescence induction in most cell culture experiments, but in vivo it is not sufficient to indicate stable cell cycle arrest and cellular senescence.

    “Thus, the lack of PCNA or KI67 staining or thymidine analog incorporation can be a strong indicator for senescence induction in most cell culture experiments, but the lack of proliferation in vivo is not sufficient to indicate stable cell cycle arrest and cellular senescence.”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Interpretationreasoning

    Auxiliary markers correspond to core senescence features including nuclear envelope erosion, reduced proliferation markers, alarmin activation, DNA damage, chromatin abnormalities, pro-secretory phenotypes, metabolic alterations, oxidative damage, and lysosomal alterations.

    “Auxiliary markers pertain to the core features of senescence, including erosion of the nuclear envelope, a decline in proliferation markers, activation of alarmins, DNA damage, chromatin abnormalities, pro-secretory phenotypes, metabolic alterations, oxidative damage, and lysosomal alterations (Fig. 1).”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S4 (based).

    Interpretationnone given

    The cell cycle inhibitors p21Cip1/Waf and/or p16Ink4a are integral to senescence both in situ and in vivo.

    “In summary, the cell cycle inhibitors p21Cip1/Waf and/or p16Ink4a are integral to senescence, both in situ and in vivo, and we recommend that every rodent study utilize at least one of these.”
    Quote in the paperNot a substantial claim
    • Rests on an assumption: p21Cip1/Waf and p16Ink4a are the cyclin-dependent kinase inhibitors most commonly expressed by senescent cells in vivo.

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Study design and conductreasoning

    The authors categorized senescence markers as primary and auxiliary based on published evidence and agreement among the research community working on the article.

    “Based on published evidence and agreement of the research community working on this article, we categorized these markers as primary and auxiliary.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Interpretationreasoning

    Numerous liver cell types display markers associated with senescence during aging, injury, regeneration, obesity, and age-related diseases, with different cell types displaying different senescence markers.

    “Numerous liver cell types, including hepatocytes, endothelial cells, hepatic stellate cells, and cholangiocytes, display markers associated with senescence during aging, injury and regeneration, obesity, and age-related diseases, with different cell types displaying different senescence markers (Fig. 2 E).”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Interpretationnone given

    These models affect only a single component of the senescence machinery and thus might not reflect the whole biology of cellular senescence.

    “While working with these models, it is important to be aware that they affect only a single component of the senescence machinery and thus might not reflect the whole biology of cellular senescence.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Premisereasoning

    Many models for senescence reporting utilize transgenes driven by different parts of the native promoter of the senescence regulatory gene, making it challenging to compare the results between models.

    “Furthermore, many models for senescence reporting utilize transgenes driven by different parts of the native promoter of the senescence regulatory gene (Table 2), making it challenging to compare the results between models.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites Table 2

      Couldn’t be matched to a paper in Crossref, so it can’t be checked.

  • May combine two claims.

    Premisereasoning

    Reporters using eGFP or tdTomato fluorescence enable the detection of p21+ or p16+ cells in situ, whereas models with luciferase enable the approximation of senescent cells in live animals by measuring light emitted after substrate administration.

    “Reporters using eGFP or tdTomato fluorescence enable the detection of p21+ or p16+ cells in situ, whereas models with luciferase enable the approximation of senescent cells in live animals by measuring the amount of light emitted after the administration of an enzymatic substrate for the luciferase reporter.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Table 2

      Couldn’t be matched to a paper in Crossref, so it can’t be checked.

  • May combine two claims.

    Interpretationreasoning

    These models are suitable for estimating the quantity of a particular type of senescent cell, but the results must be validated with additional senescence markers since single molecular endpoints are not specific for senescence.

    “While these models are suitable for estimating the quantity of a particular type of senescent cell, the results must be validated with additional senescence markers since single molecular endpoints are not specific for senescence.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Interpretationnone given

    The activity of the transgene is dependent on a single gene, which is inadequate for fully defining the senescent cell burden in vivo.

    “Similar to other transgenic models used in senescence research, the activity of the transgene is dependent on a single gene, which is inadequate for fully defining the senescent cell burden in vivo.”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Interpretationreasoning

    It is possible that the elimination of senescence in other tissues contributed to the observed phenotype.

    “Therefore, it is possible that the elimination of senescence in other tissues contributed to the observed phenotype.”
    Quote in the paperNot a substantial claim
    • Rests on an assumption: The transgene is expressed throughout the body in these models.

  • May combine two claims.

    Interpretationreasoning

    By definition, the models underestimate the senescent cell burden in vivo because they only look at a single senescent cell type and many senescence markers exhibit low expression levels, leading to low signal-to-noise ratios.

    “By definition, the models underestimate the senescent cell burden in vivo because they only look at a single senescent cell type (e.g., p16-expressing) and many senescence markers exhibit low expression levels, leading to low signal-to-noise ratios.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Interpretationreasoning

    Transcriptional profiles can approximate senescence burden, but conclusions are not definitive without orthogonal validation, ideally measuring non-mRNA endpoints.

    “Hence, although the transcriptional profile of cells or tissue regions of interest can be used to approximate the burden of senescence, the conclusions are not definitive in the absence of orthogonal validation, ideally measuring non-mRNA endpoints.”
    Quote in the paperSubstantial claim
    • Rests on an assumption: There is no evidence that cellular senescence is regulated at a transcriptional level.

  • Probably not a substantial claim: may not pass S3 (load-bearing), S4 (based).

    Interpretationreasoning

    Applying omics to assess the macromolecular composition of tissues, in principle, permits the unambiguous identification of senescent cells and their detailed characterization.

    “Applying omics to assess the macromolecular composition of tissues, in principle, permits the unambiguous identification of senescent cells and their detailed characterization.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing), S4 (based).

    Interpretationreasoning

    A SASP-heavy senescence signature may not be accurate for some subsets of immune cells.

    “Understanding the methods by which a signature is developed is also critical for identifying potential biases or limitations; for example, a SASP-heavy signature may not be accurate for some subsets of immune cells.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Interpretationreasoning

    Features such as reduced proliferation, pro-inflammatory and angiogenic factors, and resistance to apoptosis are promising markers of senescence in cancer but remain context-dependent.

    “Features such as reduced proliferation, pro-inflammatory and angiogenic factors, and resistance to apoptosis provide promising markers of senescence in cancer, but will remain context-dependent for each cancer, or even for each patient within a cohort, as very few cancers present with a homogenous mutational background.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Interpretationnone given

    Detection of senescence markers in human cancers is challenging due to universal limitations of human sample analysis.

    “However, the detection of these markers in human cancers can be challenging because of the universal limitations of human sample analysis.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing), S4 (based).

    Interpretationnone given

    Human samples provide the best source of information about how cellular senescence contributes to human physiology and pathology, relative to inbred and specified pathogen-free mice.

    “Human samples provide the best source of information about how cellular senescence contributes to human physiology and pathology, relative to inbred and specified pathogen-free mice.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S4 (based).

    Interpretationnone given

    Expression of at least three senescence markers representing different properties strongly supports that observed phenomena are related to cellular senescence.

    “Evidence of the expression of at least three senescence markers representing different properties of cellular senescence in a given tissue strongly supports that the observed phenomena are indeed related to cellular senescence.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S4 (based).

    Interpretationnone given

    As of 2024, the scientific community does not have definitive recommendations for assessing cellular senescence in human tissues, except for using large sample sizes and several stable markers.

    “As of 2024, the scientific community does not have definitive recommendations for assessing cellular senescence in human tissues, except for the suggestion of using a large sample size and investigating several markers with a preference for those that are highly stable.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S4 (based).

    Interpretationnone given

    Knowledge of the functional impact of senescent cell burden in human samples is significantly less established than in mice.

    “However, our knowledge of the functional impact of the burden of senescent cells in human samples is significantly less well established than that in mice.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Interpretationreasoning

    Human samples are often small and variable, causing problems in reaching statistical significance.

    “Often, human samples, especially those collected from live subjects, are small and low in number, and this, combined with the high interindividual variability, poses problems in reaching statistical significance.”
    Quote in the paperNot a substantial claim
    • Rests on an assumption: There is little control over variables in humans that affect senescent cell burden.

  • Probably not a substantial claim: may not pass S3 (load-bearing), S4 (based).

    Interpretationnone given

    Genetic heterogeneity between individuals likely influences the distribution of senescence biomarkers.

    “Genetic heterogeneity between individuals in the human population will likely influence the distribution of senescence biomarkers from person to person.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Interpretationnone given

    The impact of tumor cell senescence on tumor progression is not well defined and is context dependent.

    “While the impact of tumor cell senescence on tumor progression is currently not well defined, it is clearly context dependent.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Interpretationreasoning

    The diversity of cancer types and lack of established senescence markers related to cell cycle control make defining cancer cell senescence challenging.

    “The sheer diversity of cancer types (Fig. 3) and the lack of multiple established senescence markers related to cell cycle control to define cancer cell senescence in these diverse tumor types remain challenging.”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Interpretationnone given

    The present version of MICSE is based on current knowledge and will need to be updated as understanding increases.

    “Finally, the present version of the MISCE is based on current knowledge and will need to be updated as our understanding of senescence in vivo increases.”
    Quote in the paperNot a substantial claim

  • Classified as probably not a substantial claim. May combine two claims.

    Interpretationreasoning

    The majority of specific recommendations are applicable to mouse models, while human samples, non-mammalian animals, and tumors require further research before confident recommendations can be made.

    “Importantly, the majority of our specific recommendations are applicable to mouse models, as studies in human samples, non-mammalian animals, and tumors require further research before we feel confident in providing clear recommendations without being too restrictive for research progress.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Interpretationreasoning

    Senescent cells in tumors present the same challenges as those in normal tissues; they need proper identification and functional evaluation of their modulation or elimination.

    “Thus, in general, senescent cells in tumors present the same challenges as those in normal tissues; they need to be properly identified, and the impact of their modulation or elimination must be functionally evaluated in tumor tissues.”
    Quote in the paperSubstantial claim
86 of 86

  • The paper relies on this without support.

    The lack of specific and broadly applicable markers makes it difficult to identify and characterize senescent cells in tissues and living organisms.

    “However, the lack of specific and broadly applicable markers makes it difficult to identify and characterize senescent cells in tissues and living organisms.”

  • The paper relies on this without support.

    Classical senescence markers discovered and established in cell culture have limited utility for experimentation in the natural tissue microenvironment or in situ.

    “An important limitation is that classical markers of senescence, originally discovered and established using cell culture models, have limited utility for experimentation in the natural tissue microenvironment, or “in situ”.”

  • The paper relies on this without support.

    The absence of a specific cellular senescence biomarker requires measuring multiple endpoints, including nucleic acids, proteins, and enzymatic activity in the same or adjacent tissue sections.

    “In addition, the lack of a specific biomarker for cellular senescence demands that multiple endpoints be measured, requiring multiplexing and measuring nucleic acids, proteins, and enzymatic activity in the same or adjacent tissue sections.”

  • The paper relies on this without support.

    No single biomarker is sufficient to detect cellular senescence in vivo, and multiplexing remains a necessary requirement.

    “No single biomarker is sufficient to detect cellular senescence in vivo, and multiplexing remains a necessary requirement.”

  • The paper relies on this without support.

    p21Cip1/Waf and p16Ink4a are the cyclin-dependent kinase inhibitors most commonly expressed by senescent cells in vivo.

    “While there are multiple cyclin-dependent kinase inhibitors (CDKIs), p21Cip1/Waf (CDKN1A locus; encoding p21Cip1/Waf) and p16Ink4a (CDKN2A locus; encoding p16Ink4a) are the inhibitors most commonly expressed by senescent cells in vivo (Fig. 1 A).”

  • The paper relies on this without support.

    In situ skin fibroblasts are much smaller than in vitro, limiting histological visualization.

    “In contrast to in vitro conditions, skin fibroblasts in situ are much smaller and only a fraction of their volume can be visualized by histology.”

  • The paper relies on this without support.

    Keratinocytes and fibroblasts differ dramatically in replication dynamics, capacity, and lifespan in vivo.

    “since keratinocytes and fibroblasts differ dramatically in replication dynamics, capacity, and lifespan in vivo”

  • The paper relies on this without support.

    Bone is a hard tissue that requires special processing and presents methodological challenges for detecting senescence markers, particularly histology-based ones.

    “Bone is an especially hard tissue that requires special processing and presents several methodological challenges for the detection of senescence markers, particularly those that rely on histology.”

  • The paper relies on this without support.

    Bone has a specific resorption-deposition cycle and extensive endocrine regulation.

    “In addition, bone has a very specific resorption-deposition cycle and extensive endocrine regulation.”

  • The paper relies on this without support.

    Mature adipose tissue is composed of >50–90% terminally differentiated post-mitotic adipocytes, ~10% preadipocytes, and ~10% immune cells.

    “Mature adipose tissue is composed of >50–90% terminally differentiated post-mitotic adipocytes, ~10% preadipocytes, and ~10% immune cells, including macrophages and lymphocytes.”

  • The paper relies on this without support.

    SA-β-gal staining is a common method for detecting senescent cells in adipose tissue.

    “Although SA-β-gal staining is a common method for detecting senescent cells in adipose tissue”

  • The paper relies on this without support.

    Adipose tissue cellular composition varies by anatomical location, sex, and obesity, contributing to senescence heterogeneity.

    “Moreover, adipose tissue differs greatly in cellular composition based on anatomical location, sex, and degree of obesity, which undoubtedly contributes to the heterogeneity of cellular senescence.”

  • The paper relies on this without support.

    Muscle is a complex tissue composed of post-mitotic multinucleated myofibers and a heterogeneous population of mononuclear cells including satellite cells, FAPs, endothelial cells, and macrophages.

    “Muscle is a complex tissue composed of post-mitotic multinucleated cells, referred to as myofibers, and a heterogeneous population of mononuclear cells, encompassing satellite cells, fibroadipogenic progenitors (FAPs), endothelial cells, and macrophages (Fig. 2 G).”

  • The paper relies on this without support.

    Fluorescent SPiDER-ß-Gal identifies senescent-like immune cell subsets by flow cytometry with high sensitivity.

    “In addition, fluorescent SPiDER-ß-Gal has been used to identify senescent-like subsets of immune cells by flow cytometry with high sensitivity.”

  • The paper relies on this without support.

    Measurements of cellular senescence in animals and assessment of the impact of senescent cells on physiology and pathology can be aided by a wide range of transgenic mouse models.

    “Measurements of cellular senescence in animals and assessment of the impact of senescent cells on physiology and pathology can be aided by a wide range of transgenic mouse models.”

  • The paper relies on this without support.

    It is possible to drive senescence in vivo by using a variety of drugs that promote cellular, organelle, or macromolecular stress.

    “It is also possible to drive senescence in vivo by using a variety of drugs that promote cellular, organelle, or macromolecular stress.”

  • The paper relies on this without support.

    The majority of currently available models are constitutive, while inducible KO or overexpression models are in development.

    “While the majority of currently available models are constitutive (active from conception), models of inducible KO or overexpression of senescence modulators are in development.”

  • The paper relies on this without support.

    p21, p16, TP53 (p53), and other senescence-associated proteins are involved in physiology distinct from cellular senescence.

    “Moreover, p21, p16, TP53 (p53), and other senescence-associated proteins are involved in physiology that is distinct from cellular senescence.”

  • The paper relies on this without support.

    Senescence can be induced efficiently by accelerated telomere shortening, disruption of the cell division machinery, and increased DNA damage levels in vivo.

    “In accordance with research on fibroblasts in vitro, senescence can be induced efficiently by accelerated telomere shortening, disruption of the cell division machinery, and increased DNA damage levels in vivo.”

  • The paper relies on this without support.

    Genes that induce accelerated senescence upon genetic depletion play pleiotropic roles in normal physiology.

    “Genes that induce accelerated senescence upon genetic depletion play pleiotropic roles in normal physiology.”

  • The paper relies on this without support.

    Luminescence signals are strongly influenced by tissue density and light penetration, which may bias measurement of senescent cell burden.

    “Luminescence signals are strongly influenced by tissue density and light penetration, which may bias the measurement of the senescent cell burden.”

  • The paper relies on this without support.

    Reporter construct design influences the reporter signal.

    “In addition to the caveats of various reporter genes, reporter construct design also influences the reporter signal.”

  • The paper relies on this without support.

    In pharmacogenetic elimination models, an inactive apoptosis-inducing gene is expressed from a senescence regulator promoter, and drug administration eliminates p21+ or p16+ cells.

    “In these mice, an inactive form of an apoptosis-inducing gene is expressed from the promoter of a key senescence regulator, such as p21 or p16, and administration of a drug to the mice drives the elimination of p21+ or p16+ cells.”

  • The paper relies on this without support.

    These models are typically designed to enable drug-inducible elimination and to report senescence levels in situ and/or in vivo.

    “These models are typically designed to not only enable drug-inducible elimination of senescent cells, but also to report the level of cellular senescence in situ and/or in vivo.”

  • The paper relies on this without support.

    The transgene is expressed throughout the body in these models.

    “it should be acknowledged that the transgene is expressed throughout the body.”

  • The paper relies on this without support.

    Our understanding of senescence in non-mammals is limited by the shortage of reagents for molecular analysis in many of these organisms.

    “However, our understanding of senescence in non-mammals is limited by the shortage of reagents for molecular analysis in many of these organisms.”

  • The paper relies on this without support.

    Cells from non-mammals are capable of undergoing cell cycle arrest resembling cellular senescence, but it might be driven by alternative mechanisms.

    “Although cells from non-mammals are capable of undergoing cell cycle arrest that resembles cellular senescence, it might be driven by alternative mechanisms.”

  • The paper relies on this without support.

    Other markers of cellular senescence, such as SA-β-gal, appear to function similarly across different animals, and their use has been instrumental in determining the presence of senescence in vivo across organisms.

    “Other markers of cellular senescence, such as SA-β-gal, appear to function similarly across different animals, and their use has been instrumental in determining the presence of senescence in vivo across organisms.”

  • The paper relies on this without support.

    The impact of senescent cells on non-mammalian biology has been mainly investigated in development, tissue remodeling, and regeneration, while disease and aging need further exploration.

    “The impact of senescent cells on the biology of non-mammalian species has been mainly investigated in the context of development, tissue remodeling, and regeneration, while conditions such as disease and aging still need to be further explored.”

  • The paper relies on this without support.

    The omics revolution enables in-depth characterization of the macromolecular composition of biological samples.

    “The omics revolution enables in-depth characterization of the macromolecular composition of biological samples.”

  • The paper relies on this without support.

    Senescent fibroblasts produced in vitro have been thoroughly characterized using epigenomic, transcriptomic, proteomic, lipidomic, metabolomic, and phenomic approaches.

    “Senescent fibroblasts produced in vitro have been thoroughly characterized using epigenomic, transcriptomic, proteomic, lipidomic, metabolomic, and phenomic approaches.”

  • The paper relies on this without support.

    Spatial omics platforms allow correlation of an omic dataset with spatial location within a tissue and have the potential to revolutionize in vivo senescence research.

    “Furthermore, spatial omics platforms allow for the correlation of an omic dataset (currently either whole genome transcriptomics or targeted transcriptomics or proteomics) with spatial location within a tissue and have the potential to revolutionize in vivo senescence research.”

  • The paper relies on this without support.

    Both bulk and single-cell omics have been applied to in vivo senescence research; however, analysis and interpretation pose challenges.

    “Both bulk and single-cell omics have been applied to in vivo senescence research; however, the analysis and interpretation of results pose challenges.”

  • The paper relies on this without support.

    Spatial transcriptomics is less sensitive than scRNA-seq and can introduce bias due to detection methods that do not always distinguish splice variants of CDKI genes such as CDKN2A.

    “Spatial transcriptomics is less sensitive than scRNA-seq and can introduce bias based on methods of either detection of the 3’ end of (polyA tail) or nucleic acid ISH probes, which do not always distinguish between splice variants of CDKI genes such as CDKN2A.”

  • The paper relies on this without support.

    Single-cell transcriptomics requires a large number of intact cells.

    “Single-cell transcriptomics requires a large number of intact cells.”

  • The paper relies on this without support.

    Multiple sources of bias can be introduced through tissue dissociation, microfluidics, data analysis, and statistics.

    “Furthermore, multiple sources of bias can be introduced through tissue dissociation (loss of fragile cells), microfluidics (loss of large cells), data analysis, and statistics.”

  • The paper relies on this without support.

    There is no evidence that cellular senescence is regulated at a transcriptional level.

    “Finally, there is no evidence that cellular senescence is regulated at a transcriptional level.”

  • The paper relies on this without support.

    Changes in nuclear and cellular morphology are largely conserved across senescent cells in vitro and in vivo.

    “Changes in nuclear and cellular morphology are largely conserved across senescent cells, both in vitro and in vivo.”

  • The paper relies on this without support.

    Methods for detecting anti-apoptotic proteins in situ or in vivo are not yet well established.

    “Methods for detecting anti-apoptotic proteins in situ or in vivo are not yet well established but would provide targetable senescence biomarkers in the context of cancer.”

  • The paper relies on this without support.

    p16 is easier to detect in humans than in murine samples due to a reliable antibody against human p16Ink4a.

    “Some markers of senescence, such as p16, are easier to detect in humans than in murine samples (because of the availability of a reliable antibody against human p16Ink4a developed as a clinical prognostic tool for cancer).”

  • The paper relies on this without support.

    Metabolically active and lipid-rich tissues are more vulnerable to rapid ex vivo degradation.

    “Similarly, metabolically active and lipid-rich tissues are more vulnerable to rapid ex vivo degradation.”

  • The paper relies on this without support.

    Aging drives heterogeneity among individuals, which has not been observed in young individuals.

    “Aging also drives incredible heterogeneity among individuals, which has not been observed in young individuals.”

  • The paper relies on this without support.

    There is little control over variables in humans that affect senescent cell burden.

    “Furthermore, unlike laboratory models, there is little control over variables in humans that affect senescent cell burden, such as occult disease, body mass index, environmental factors, genetic factors impacting genome instability and telomere length, and levels of psychosocial stress.”

  • The paper relies on this without support.

    There is currently limited knowledge about SASP factors in liquid biopsies.

    “While surrogate biomarkers of senescence burden from liquid biopsies, preferably from minimally invasive sampling such as blood, saliva, wound pus, urine, synovial, or cerebrospinal fluid, would provide a necessary and more accessible alternative, there is currently limited knowledge about SASP factors in these liquid biopsies.”
44 of 44

Claims other papers make about this one

Each is checked against this paper’s own text: does it say what the citing paper says it does?

  • Aging biology differs between rodents and humans, and findings from a mouse model may not fully translate to human liver.

    “aging biology differs between rodents and humans, and findings from a mouse model may not fully translate to human liver [5,51,52].”

    From Fluorescence-Guided Spatial Mapping of p21-Expressing Senescent Cells in Aged Liver at Single-Cell Resolution (2026) · cites this paper as [5]

    The “Questions to be answered” section (page unavailable) says rats model human physiology and many aspects of aging better than mice, supporting caution about translating mouse findings to humans. The “Liver” section describes senescence findings in murine liver, but does not compare them with human liver or specifically assess whether those findings translate. (LLM, conf=0.82)

    Experimental — accuracy not yet measured

  • Senescent cells are typically rare, heterogeneous, and context dependent, causing difficulties in identifying them and defining their tissue-level interactions.

    “A persistent challenge is that senescent cells are typically rare, heterogeneous, and context dependent, causing difficulties in identifying them and defining their tissue-level interactions [5].”

    From Fluorescence-Guided Spatial Mapping of p21-Expressing Senescent Cells in Aged Liver at Single-Cell Resolution (2026) · cites this paper as [5]

    The Heterogeneity section says senescence phenotypes vary by organ, cell type, and stimulus, making a single marker inadequate for identification. Challenges and limitations says senescent cells are often estimated at under 10% and that studying these rare cells requires demanding single-cell or spatial methods. The passages do not specifically establish that these features make tissue-level interactions difficult to define. (LLM, conf=0.83)

    Experimental — accuracy not yet measured

  • No individual marker works universally to detect senescent cells in every context.

    “our findings reinforce the notion that no single marker can universally identify senescent cells across all contexts, as previously proposed,1,7”

    From SenCat: Cataloging human cell senescence through multi-omic profiling of multiple senescent primary cell types (2026) · cites this paper as [7]

  • Current guidelines by the senescence research community prioritize p21 and p16 as key CDKi for senescence characterization.

    “Yet, current guidelines by the senescence research community prioritize p21 and p16 as key CDKi for senescence characterization (Ogrodnik et al, 2024; Suryadevara et al, 2024).”

    From Distinct senotypes in p16- and p21-positive cells across human and mouse aging tissues (2025) · cites this paper as Ogrodnik et al, 2024

  • The study utilized multiple complementary methods to characterize and confirm a senescence phenotype following demyelination, in line with recently released guidance for studying senescence.

    “With this in mind, here we utilized multiple complementary methods to characterize and confirm a senescence phenotype following demyelination, in line with recently released guidance for studying senescence76.”

    From Senescent-like microglia limit remyelination through the senescence associated secretory phenotype (2025) · cites this paper as [76]

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

Each contradiction between two passages listed here was confirmed by an independent check.

  • The acronym is written two different ways

    Minor

    The document introduces the guideline set as MICSE, but later refers to the same guideline name as MISCE. Those two spellings cannot both be the correct acronym for the same project.

1 of 1

Cites

Flagged first, then by citations

1 of the 170 references couldn’t be matched to a known paper.

Cited by

Sorted by citations

Related

Authors
Ogrodnik, M., Carlos Acosta, J., Adams, P. D., d’Adda di Fagagna, F., Baker, D. J., Bishop, C. L., Chandra, T., Collado, M., Gil, J., Gorgoulis, V., Gruber, F., Hara, E., Jansen-Dürr, P., Jurk, D., Khosla, S., Kirkland, J. L., Krizhanovsky, V., Minamino, T., Niedernhofer, L. J., Passos, J. F., Ring, N. A. R., Redl, H., Robbins, P. D., Rodier, F., Scharffetter-Kochanek, K., Sedivy, J. M., Sikora, E., Witwer, K., von Zglinicki, T., Yun, M. H., Grillari, J., & Demaria, M.
Affiliations
  • Ludwig Boltzmann Gesellschaft (AT)
  • Sanford Burnham Prebys Medical Discovery Institute (US)
  • Mayo Clinic (US)
  • Queen Mary University of London (GB)
  • University of Edinburgh (GB)
  • Consejo Superior de Investigaciones Científicas (ES)
  • University of Dundee (GB)
  • Immunology Frontier Research Center, The University of Osaka (JP)
  • Weizmann Institute of Science (IL)
  • Japan Agency for Medical Research and Development (JP)
  • University of Minnesota (US)
  • Brown University (US)
  • Instytut Biologii Doświadczalnej im. Marcelego Nenckiego (PL)
  • Johns Hopkins University (US)
  • Max Planck Institute of Molecular Cell Biology and Genetics (DE)
  • Institute of Molecular Biotechnology (AT)
  • University Medical Center Groningen (NL)
Corresponding author
  • Ogrodnik, Mikolaj
  • Grillari, Johannes
  • Demaria, Marco
ORCID
Venue
Cell
Publisher
Elsevier BV
Published
2024-08
Volume
187
Issue
16
Pages
4150-4175
ISSN
0092-8674 (ISSN-L)
DOI
10.1016/j.cell.2024.05.059
PMC
PMC11790242
OpenAlex
W4401420229
Cited by
376
References
170
Citations by year
2026: 206 · 2025: 142 · 2024: 27 · 2023: 1
FWCI
21.30
Type
journal article
Language
en
Open access
Licence
Locations
  • Cell (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • PubMed Central (acceptedVersion)
  • Queen Mary Research Online (Queen Mary University of London) (submittedVersion, cc-by)
  • UCrea (University of Cantabria) (submittedVersion, cc-by)
  • DIGITAL.CSIC (Spanish National Research Council (CSIC)) (submittedVersion, cc-by)
  • University of Groningen research database (University of Groningen / Centre for Information Technology) (submittedVersion, cc-by)
  • Digital Library of the University of Innsbruck (University of Innsbruck) (submittedVersion, cc-by)
  • Newcastle University ePrints (Newcastle Univesity) (cc-by)
Funders
  • Glenn Foundation for Medical Research
  • Pfizer
  • Brown University
  • Cancer Research Society (20087)
  • Merck KGaA
  • Buck Institute for Research on Aging
  • Noaber Foundation
  • Ovarian Cancer Canada
  • Cancer Research UK (C15075/A28647)
  • Imperial College London (H2020)
  • European Commission (2014‐2020, HORIZON2020, H2020, 857524, POR FESR 2014-2020, 825575, 70/3/8916, 856487)
  • Deutsche Forschungsgemeinschaft (450807335, 825575, GRK-1789, 450627322, CRC 1506, 497658823, 22137416)
  • ZonMw
  • Austrian Science Fund (P36483, P 35268, P 37321, P35382)
  • Fondation pour la Recherche Médicale
  • Nederlandse Organisatie voor Wetenschappelijk Onderzoek (09150172010029)
  • General Secretariat for Research and Technology (2020SE01300001, H2020)
  • Herzfelder'sche Familienstiftung
  • Associazione Italiana per la Ricerca sul Cancro (21762, 21091)
  • Universität für Bodenkultur Wien
  • Allgemeine Unfallversicherungsanstalt
  • Zentrum für Regenerative Therapien Dresden
  • Regione Lombardia (POR FESR 20142020, 2014–2020, FESR 2014-2020)
  • Ludwig Boltzmann Gesellschaft
  • Fondazione Regionale per la Ricerca Biomedica (825575)
  • National Institutes of Health (ASAP-000592, H2020, AG031862)
  • Horizon 2020 Framework Programme (825575, 857524)
  • Natural Sciences and Engineering Research Council of Canada
  • Fonds de Recherche du Québec - Santé (281706, 33070, 22624)
  • Medical Research Council (MC-U120085810)
  • Biotechnology and Biological Sciences Research Council (BB/S006710/1)
  • European Regional Development Fund (FESR 2014-2020, 2014-2020’, H2020, POR FESR 2014-2020)
  • Agencia Estatal de Investigación (H2020)
  • National Institute on Aging (P01AG031862, P01AG043376, R01AG068182, R01AG053229, R01AG068076, R01AG068048, R33AG061456, R37AG013925, R01AG082708, U19AG056278, U54AG079779, U54AG079754, U54AG076041, P01AG062413, U54AG079758, R01AG071861, P01AG051449, R01AG063543)
  • National Cancer Institute (UH3CA268103, UG3CA268103)
Topics
  • Health Sciences › Medicine › Physiology › Telomeres, Telomerase, and Senescence
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Aging › Genetics, Aging, and Longevity in Model Organisms
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Single-cell and spatial transcriptomics
Keywords
cellular senescence, senescence markers, senolytics, cell cycle arrest, senescence-associated secretory phenotype, tumors, reporting guidelines
MeSH terms
Animals, Humans, Neoplasms, Biomarkers, Cellular Senescence, Guidelines as Topic

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Claims read by deepseek-v4.1-flash under definition version 3 (extract-v3) on 2026-09-28.

Classified by jev-1.13-20260917 (gold-guideline-v0.2) on 2026-09-28.

Cited sources are checked by gpt-6-sol.

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

What counts as a claim ›How we measure confidence ›How we check this ›

Are you an author, or do you see a problem with this page? Request a review